CN117099612A - Cordyceps militaris cultivation and purification method - Google Patents
Cordyceps militaris cultivation and purification method Download PDFInfo
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- CN117099612A CN117099612A CN202311127456.XA CN202311127456A CN117099612A CN 117099612 A CN117099612 A CN 117099612A CN 202311127456 A CN202311127456 A CN 202311127456A CN 117099612 A CN117099612 A CN 117099612A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/20—Culture media, e.g. compost
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/60—Cultivation rooms; Equipment therefor
- A01G18/69—Arrangements for managing the environment, e.g. sprinklers
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Mycology (AREA)
- Environmental Sciences (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention provides a cordyceps militaris cultivation and purification method, and belongs to the technical field of cultivation; the cultivation method is based on cultivation equipment; the incubation apparatus includes: the device comprises a culture assembly, a soaking assembly, a detection assembly and a pupa formation assembly, wherein the soaking assembly is arranged on one side of the interior of the culture assembly, the detection assembly is arranged at the bottom of the culture assembly, and the pupa formation assembly is arranged on the other side of the interior of the culture assembly; the cultivation method comprises the following steps: material preparation: and (3) putting the larvae into the culture assembly, adding needed nutrient solution and flavoring agent into the soaking assembly, putting the leaf ends of the culture leaves into the culture assembly, and providing nutrients with the nutrient solution and the flavoring agent for the larvae. In conclusion, the invention has the advantages of reducing the rejection rate of directly adding the nutrient solution and the flavoring agent into the cordyceps militaris, improving the nutrition content of the cultivated cordyceps militaris, controlling the taste of the cordyceps militaris, further improving the purity of cordycepin and the like.
Description
Technical Field
The invention relates to the technical field of cultivation, in particular to a cultivation and purification method of cordyceps militaris.
Background
Cordyceps militaris is also called Cordyceps militaris, and belongs to the order of Clavipitaceae, belonging to the genus Cordyceps. Is widely distributed around the world, and has growth in Jilin, hebei, sichuan, hunan, hubei, shanxi provinces and the like in China. The folk medicine is used for treating diseases such as pneumonia, kidney deficiency, lumbago and the like. The requirements of the cordyceps militaris on the growth environment are low, mycelium pellets can be formed by liquid fermentation, and a sub-base can be obtained by artificial large-scale solid culture. Cordyceps militaris contains phosphorus, potassium, magnesium and other inorganic elements, proteins, amino acids, saccharides, fat, alkaloid volatile oil, cordycepic acid, cordycepin, superoxide dismutase (SOD), nucleotides and other organic matters. Among them, cordycepin (3' -deoxyadenosine) is also called cordycepin and cordycepin is one of the important effective components.
At present, the nutrition components of the cordyceps militaris are basically fixed, and how to further improve the cordycepin content of the artificially cultivated cordyceps militaris is a difficult problem, because in the strain cultivation link in the process of cultivating the cordyceps militaris, the cordyceps militaris parasitical pupa is cultivated firstly, and a nutrient solution beneficial to the growth of the cordyceps militaris is provided for the cordyceps militaris. Most of the existing nutrient solution adding modes are needle tube injection and nutrient solution soaking, however, in the needle tube injection mode, the nutrient solution is poor in absorption effect of the pupa, and because the nutrient solution is directly injected into the pupa, compared with the normal state, the cordyceps militaris and nutrients are easy to repel due to lack of conversion and absorption links of the pupa, nutrient substance unbalance in a certain aspect is also easy to be caused, and the quality and the survival rate of the pupa are reduced; under the mode of soaking the nutrient solution, the nutrient solution is positioned on the outer surface of the pupa, so that the absorption effect of the pupa on the nutrient solution is very limited, parasitic help to the cordyceps militaris is small, the concentration of part of the nutrient solution is higher than that of the cell sap in the pupa, at the moment, water can flow from low concentration to high concentration, the process is called osmosis, if the osmosis is too severe, normal functions of the pupa cells are lost and finally the pupa cells die, and therefore, how to cultivate the pupa with higher quality and survival rate is a difficult problem. Meanwhile, after the cordyceps militaris is ripe, the cordyceps militaris has light fishy smell after being eaten, and part of people are difficult to adapt to the taste of the cordyceps militaris, so that the audience surface of the cordyceps militaris is reduced, and how to customize and improve the taste of the cordyceps militaris is a difficult problem.
Therefore, the invention needs a method for cultivating and purifying the cordyceps militaris, which further improves the cordycepin content of the artificially cultivated cordyceps militaris and solves the problem of reduced audience surface of the cordyceps militaris caused by the fishy smell of the cordyceps militaris.
Disclosure of Invention
The invention aims to solve the technical problem of providing a cultivation and purification method of cordyceps militaris, which reduces the rejection rate of directly adding nutrient solution and flavoring agent into pupa through material preparation in the cultivation method so as to improve the nutrition content of cultivated cordyceps militaris and control the taste of cordyceps militaris; through the cooperation setting of extraction element and purification subassembly, have the advantage such as further improvement cordycepin's purity of extracting.
In order to solve the technical problems, the invention provides the following technical scheme:
a method for cultivating cordyceps militaris,
the cultivation method is based on cultivation equipment;
the incubation apparatus includes: the device comprises a culture assembly, a soaking assembly, a detection assembly and a pupa formation assembly, wherein the soaking assembly is arranged on one side of the interior of the culture assembly, the detection assembly is arranged at the bottom of the culture assembly, the pupa formation assembly is arranged on the other side of the interior of the culture assembly, the soaking assembly comprises a soaking box, a liquid inlet and a water outlet pipe, the liquid inlet is arranged at the top of the soaking box, and the water outlet pipe is arranged on one side of the soaking box;
The soaking tank comprises a tank body, a water storage space, a motor, stirring blades, a guide sponge layer and a water storage sponge layer, wherein the water storage space is formed in the bottom of the inner wall of the tank body and is communicated with the liquid inlet, the motor is arranged at the bottom of the tank body, the stirring blades are arranged at the bottom of the water storage space and are connected with the motor, the top of the inner wall of the tank body is filled with the water storage sponge layer, the guide sponge layer is arranged between the water storage space and the water storage sponge layer, and the water storage sponge layer is communicated with the water outlet pipe;
the cultivation method comprises the following steps:
material preparation: placing the larvae into the culture assembly, adding absorption liquid into the soaking assembly, inserting stem ends of the culture leaves into the soaking assembly, placing leaf ends of the culture leaves into the culture assembly, and providing nutrients with the absorption liquid for the larvae;
preparing environment: detecting the state of the larvae through the detection assembly, keeping the increase of the defecation amount of the larvae stable, picking out unqualified larvae, and waiting for the qualified larvae to form pupae in the pupae formation assembly;
seed selection of strains: selecting excellent strains, sterilizing and extracting strains;
Culturing strains: sterilizing the surface of the pupa, infecting the pupa by strain, and culturing the infected pupa in aseptic environment.
Preferably, the culture assembly comprises a culture bottom plate, filtering holes, coamings, a rotating shaft and a movable shell, wherein the surface of the culture bottom plate is provided with the filtering holes in an array, the coamings are arranged on the periphery of the top of the culture bottom plate, the rotating shafts are arranged on two sides of the outer wall of the coamings, the movable shell is arranged on one side of the rotating shaft, and the movable shell is sleeved outside the culture bottom plate.
Preferably, the soaking assembly further comprises a valve, clamping plates, tension springs and a connecting block, the soaking box is detachably arranged on one side of the top of the culture bottom plate, the inlet end of the water outlet pipe is communicated with the soaking box, the outlet end of the water outlet pipe is provided with the valve, the valve is used for enabling liquid inside the soaking box to flow unidirectionally from the inlet end of the water outlet pipe to the outlet end, the clamping plates are arranged on one side of the water outlet pipe away from the soaking box, the number of the clamping plates is two, the tension springs are arranged between the two clamping plates, and the connecting block is arranged on one side of the soaking box close to the coaming.
Preferably, the detection assembly comprises a collecting box, a display screen, a weight detector, an outlet channel, a baffle and a sampling box, wherein the collecting box is arranged at the bottom of the culture bottom plate, the inlet end of the collecting box is communicated with the outlet end of the filtering hole, one side of the outer surface of the collecting box is provided with the display screen, the bottom of the inner wall of the collecting box is obliquely arranged, the weight detector is arranged at the bottom of the inner wall of the collecting box, the outlet channel is arranged at the other side of the outer surface of the collecting box, the baffle is movably arranged at one end of the outlet channel, and the sampling box is arranged at the outlet end of the outlet channel.
Preferably, the pupa formation assembly comprises a pupa formation outer box, a pupa formation inner box and a containing box, wherein the pupa formation outer box is arranged on the other side of the top of the culture bottom plate, the pupa formation inner box is arranged in the pupa formation outer box, and the containing box is arranged on the surface of the pupa formation inner box in an array.
Preferably, in the preparation of the material, the absorption liquid comprises a nutrient solution and a flavoring agent,
the nutrient solution comprises 65-75 parts by weight of water, glucose, sucrose, maltose, starch and pectin: 10 to 15: 2-10: 2-10: 3 to 12:0.2 to 1;
The flavoring agent comprises salty agent, sweetener, flavoring agent, sour agent and Xin Xiangji, wherein the flavoring agent comprises sodium glutamate and inosinic acid, and the ratio of the sodium glutamate to the inosinic acid is 5-20: 1, a step of;
the cultured leaf is one or more of green grass, mulberry leaf, locust tree leaf, banyan leaf, camphor leaf, willow leaf and tender leaf;
the stem end of the culture leaf is inserted into the water outlet pipe in the preparation of the material, and the leaf end of the culture leaf is placed on the culture bottom plate.
Preferably, the qualification criteria for larvae in the environmental preparation are: the larvae had no spots on the outer surface, the length of the larvae was 5 to 7cm, and the ratio of defecation amount to days of culture was 0.1g:1day, chlorophyll content in the stool is less than 15%.
The invention also provides a method for purifying Cordyceps militaris,
the purification method is based on a purification device;
the purification apparatus includes: the output end of the extraction component is connected with the input end of the purification component;
the purification method comprises the following steps:
crushing and drying: crushing the cultivated cordyceps militaris to a diameter of 150-200 um, and putting the cordyceps militaris into a baking oven with the temperature of 35-45 ℃ for baking;
filtering Cordyceps militaris powder: placing the dried Cordyceps militaris into the extraction component, pouring distilled water, heating for 10min, extracting for 30min, and filtering with 0.45 μm microporous membrane;
Purifying cordycepin: extracting cordycepin by the purification component, wherein the extraction temperature is 100 ℃, and the feed-liquid ratio is 1:15.
preferably, the extraction assembly comprises: the device comprises a filter paper piece, an extractor, a collector and an oil bath pot, wherein the filter paper piece is arranged in the extractor, the collector is connected to the bottom of the extractor, and the oil bath pot is arranged at the bottom of the collector.
Preferably, the purification assembly comprises: distiller, condenser pipe, recovery bottle and magnetic stirrer, the distiller is used for connecting the collector, one side of distiller is equipped with the condenser pipe, the one end of condenser pipe is connected in the recovery bottle, the bottom of recovery bottle is equipped with magnetic stirrer.
Compared with the prior art, the invention has at least the following beneficial effects:
according to the scheme, the nutrient solution and the flavoring agent in the soaking component are absorbed by the stem end of the culture leaf through material preparation in the cultivation method, so that the nutrient solution and the flavoring agent are fused into the leaf end of the culture leaf, therefore, after the culture leaf containing personnel-controllable microelements is eaten by the larva, the larva can absorb the nutrient solution and the flavoring agent into the body in a conversion absorption mode, the nutrient solution and the flavoring agent are converted into micromolecular substances after being digested and absorbed by the larva, the nutrient components and the required flavoring agent contained in the larva are gradually improved and increased in a long-term adult period, the rejection rate of directly adding the nutrient solution and the flavoring agent into the pupa is reduced, the nutrient solution and the flavoring agent are accumulated in the body of the larva in a conversion absorption mode, the nutrient solution and the flavoring agent are absorbed by infected strain, the nutrient content of the cultivated cordyceps militaris is improved, the taste of the cordyceps militaris is controlled through the residual flavoring agent, the customization of the taste is convenient, more people adapt to the cordyceps militaris, and the audience face and the prospect of the cordyceps militaris is expanded.
Through the combination setting of soaking case, can utilize stirring leaf in retaining space bottom to stir nutrient solution and the flavouring of injection continually for nutrient solution and flavouring intensive mixing, and prevent that the element of nutrient solution and flavouring from depositing, simultaneously, through guiding sponge layer and water storage sponge layer, make the stem tip of cultivateing the leaf contact with water storage sponge layer, make the stem tip of cultivateing the leaf receive the water storage sponge layer parcel that has absorbed nutrient solution and flavouring, make the root and the surface of the stem tip of cultivateing the leaf all contact with water storage sponge layer, the area of contact of the stem tip of cultivateing the leaf and water storage sponge layer has been improved, increase the absorption efficiency of the stem tip of cultivateing the leaf, and after the water level of nutrient solution and flavouring descends in retaining space, through guiding the conduction of sponge layer, the stem tip of cultivateing the leaf still can be stable absorb nutrient solution and flavouring, keep the sufficient of the leaf tip nutrition of cultivateing the leaf, do not need to inject nutrient solution and flavouring continually, be convenient for use.
Through the environmental preparation in the cultivation method, the defecation amount of the larvae can be monitored in time by utilizing the weight detector, so that the defecation amount of the larvae and the cultivation days are in a normal proportion, and meanwhile, the defecation of the larvae is extracted, so that the chlorophyll content in the defecation of the larvae is in a normal value, and the absorption degree of the larvae to nutrient solution and flavoring agent is detected.
Through the cooperation setting of extraction element and purification subassembly, can carry out the purification of a lot of to the Cordyceps militaris, further improve the purification degree of cordycepin.
In conclusion, the invention has the advantages of reducing the rejection rate of directly adding the nutrient solution and the flavoring agent into the cordyceps militaris, improving the nutrition content of the cultivated cordyceps militaris, controlling the taste of the cordyceps militaris, further improving the purity of cordycepin and the like.
Drawings
The accompanying drawings, which are incorporated herein and form a part of the specification, illustrate embodiments of the present disclosure and, together with the description, further serve to explain the principles of the disclosure and to enable a person skilled in the pertinent art to make and use the disclosure.
FIG. 1 is a schematic diagram of the overall structure of cultivation equipment in a cultivation and purification method of Cordyceps militaris;
FIG. 2 is a schematic structural diagram of a culturing component in a Cordyceps militaris culturing and purifying method;
FIG. 3 is a schematic diagram of the structure of the soaking assembly in the cultivation and purification method of Cordyceps militaris;
FIG. 4 is a schematic cross-sectional structure of a detection assembly in a cordyceps militaris cultivation and purification method;
FIG. 5 is a schematic cross-sectional structure of a purifying apparatus in the cultivation and purification method of Cordyceps militaris;
FIG. 6 is a flow chart of a cultivation method in a cultivation and purification method of Cordyceps militaris;
FIG. 7 is a flow chart of a purification method in the cultivation and purification method of Cordyceps militaris;
FIG. 8 is a schematic cross-sectional structure of a soaking module in the cultivation and purification method of Cordyceps militaris.
[ reference numerals ]
1. A cultivation device; 11. a culturing assembly; 111. a culture bottom plate; 112. a filter hole; 113. coaming plate; 114. a rotating shaft; 115. a movable case; 12. a soaking component; 121. a soaking box; 1211. a case; 1212. a water storage space; 1213. a motor; 1214. stirring the leaves; 1215. guiding the sponge layer; 1216. a water storage sponge layer; 122. a liquid inlet; 123. a water outlet pipe; 124. a valve; 125. a clamping plate; 126. a tension spring; 127. a connecting block; 13. a detection assembly; 131. a collection box; 132. a display screen; 133. a weight detector; 134. an outlet channel; 135. a baffle; 136. a sampling box; 14. a pupa formation component; 141. a pupa formation outer box; 142. a pupa formation inner box; 143. a container; 2. a purifying device; 21. an extraction assembly; 211. a filter paper member; 212. an extractor; 213. a collector; 214. an oil bath pan; 22. a purification assembly; 221. a distiller; 222. a condensing tube; 223. recovering the bottle; 224. magnetic stirrer.
While particular structures and devices are shown in the drawings to enable a clear implementation of embodiments of the invention, this is for illustrative purposes only and is not intended to limit the invention to the particular structures, devices and environments, which may be modified or adapted by those of ordinary skill in the art, as desired, and which remain within the scope of the appended claims.
Detailed Description
The method for cultivating and purifying the cordyceps militaris provided by the invention is described in detail below with reference to the accompanying drawings and specific embodiments. While the invention has been described herein in terms of the preferred and preferred embodiments, the following embodiments are intended to be more illustrative, and may be implemented in many alternative ways as will occur to those of skill in the art; and the accompanying drawings are only for the purpose of describing the embodiments more specifically and are not intended to limit the invention specifically.
It should be noted that references in the specification to "one embodiment," "an example embodiment," "some embodiments," etc., indicate that the embodiment described may include a particular feature, structure, or characteristic, but every embodiment may not necessarily include the particular feature, structure, or characteristic. Further, when a particular feature, structure, or characteristic is described in connection with an embodiment, it is submitted that it is within the knowledge of one skilled in the relevant art to effect such feature, structure, or characteristic in connection with other embodiments whether or not explicitly described.
Generally, the terminology may be understood, at least in part, from the use of context. For example, the term "one or more" as used herein may be used to describe any feature, structure, or characteristic in a singular sense, or may be used to describe a combination of features, structures, or characteristics in a plural sense, depending at least in part on the context. In addition, the term "based on" may be understood as not necessarily intended to convey an exclusive set of factors, but may instead, depending at least in part on the context, allow for other factors that are not necessarily explicitly described.
It will be understood that the meanings of "on … …", "over … …" and "over … …" in this disclosure should be interpreted in the broadest sense so that "on … …" means not only "directly on" but also includes meaning "directly on" something with intervening features or layers therebetween, and "over … …" or "over … …" means not only "on" or "over" something, but also may include its meaning "on" or "over" something without intervening features or layers therebetween.
Furthermore, spatially relative terms such as "under …," "under …," "lower," "above …," "upper," and the like may be used herein for ease of description to describe one element or feature's relationship to another element or feature as illustrated in the figures. Spatially relative terms are intended to encompass different orientations of the device in use or operation in addition to the orientation depicted in the figures. The device may be otherwise oriented and the spatially relative descriptors used herein may similarly be interpreted accordingly.
As shown in fig. 1 to 7, an embodiment of the present invention provides a cultivation method of cordyceps militaris, which is based on a cultivation apparatus 1;
the incubation apparatus 1 includes: the device comprises a culture assembly 11, a soaking assembly 12, a detection assembly 13 and a pupa formation assembly 14, wherein the soaking assembly 12 is arranged on one side of the interior of the culture assembly 11, the detection assembly 13 is arranged at the bottom of the culture assembly 11, the pupa formation assembly 14 is arranged on the other side of the interior of the culture assembly 11, the soaking assembly 12 comprises a soaking box 121, a liquid inlet 122 and a water outlet pipe 123, the liquid inlet 122 is arranged at the top of the soaking box 121, and the water outlet pipe 123 is arranged on one side of the soaking box 121;
the soaking tank 121 comprises a tank body 1211, a water storage space 1212, a motor 1213, stirring blades 1214, a guiding sponge layer 1215 and a water storage sponge layer 1216, wherein the water storage space 1212 is arranged at the bottom of the inner wall of the tank body 1211, the water storage space 1212 is communicated with the liquid inlet 122, the motor 1213 is arranged at the bottom of the tank body 1211, the stirring blades 1214 are arranged at the bottom of the water storage space 1212, the stirring blades 1214 are connected with the motor 1213, the top of the inner wall of the tank body 1211 is filled with the water storage sponge layer 1216, the guiding sponge layer 1215 is arranged between the water storage space 1212 and the water storage sponge layer 1216, and the water storage sponge layer 1216 is communicated with the water outlet pipe 123;
Through the combination setting of soaking case 121, can utilize stirring leaf 1214 of retaining space 1212 bottom to stir nutrient solution and the flavouring of injection constantly for nutrient solution and flavouring intensive mixing, and prevent the element precipitation of nutrient solution and flavouring, simultaneously, through guiding sponge layer 1215 and water storage sponge layer 1216, make the stem tip of cultivateing the leaf contact with water storage sponge layer 1216, make the stem tip of cultivateing the leaf receive the water storage sponge layer 1216 parcel of absorbed nutrient solution and flavouring, make the root and the surface of the stem tip of cultivateing the leaf all contact with water storage sponge layer 1216, the area of contact of the stem tip of cultivateing the leaf and water storage sponge layer 1216 has been improved, increase the absorption efficiency of the stem tip of cultivateing the leaf, and after the water level of nutrient solution and flavouring descends in retaining space 1212, through the conduction of guide sponge layer 1215, the stem tip of cultivateing the leaf still can stable absorption nutrient solution and flavouring, keep the sufficient of the leaf tip nutrition of cultivateing the leaf, do not need to inject the nutrient solution and flavouring constantly, the use is convenient.
Note that the stirring blade 1214 is connected to the motor 1213 through a bearing housing, and a waterproof treatment is performed between the bearing housing and the case 1211.
The cultivation method comprises the following steps:
Material preparation: putting the larvae into a culture assembly 11, adding required nutrient solution and flavoring agent into a soaking assembly 12, inserting stem ends of the culture leaves into the soaking assembly 12, putting leaf ends of the culture leaves into the culture assembly 11, and providing nutrients with the nutrient solution and the flavoring agent for the larvae; it is noted that the cultured leaves are rich in chlorophyll, and the fresh leaves contain 0.2 to 0.3 percent, and the larvae basically do not absorb after feeding, so that the absorption degree of the larvae on nutrient solution and flavoring agents can be judged by observing the chlorophyll content in the excrement of the larvae.
Preparing environment: detecting the state of the larvae through the detection assembly 13, keeping the increase of the defecation amount of the larvae stable, picking out unqualified larvae, and waiting for the qualified larvae to form pupae in the pupae forming assembly 14; alternatively, the body colour of the larvae is observed: the body color of the larvae should be bright and uniform without obvious spots, discoloration or blackening. If the larvae are found to be abnormal in body color, it may mean that the larvae are ill, as a result of timely inspection. The behaviour of the larvae can also be observed: the larvae should behave actively, normally, without abnormal behavior. If abnormal behavior of the larvae is found, such as stopping feeding, continuous peristalsis, or convulsions, etc., it may mean that the silkworm suffers from diseases.
Seed selection of strains: selecting excellent strains, sterilizing and extracting strains;
culturing strains: the body surface of the insect pupa is disinfected, the insect pupa is infected by the strain, the infected insect pupa is put into a sterile environment for culture, and the insect pupa is worth noting that the insect pupa contains rich protein, fat, carbohydrate and the like, but the necessary nutrient solution such as glucose, sucrose, maltose, starch, pectin and the like still needs to be manually injected, wherein the utilization effect of the glucose, sucrose and other small molecular saccharides is the best. The nitrogen source nitrogen element is organic nitrogen such as protein and nucleic acid synthesized by Cordyceps militaris, and inorganic nitrogen such as ammonium salt. The available organic nitrogen is a lot, such as amino acid, peptone, bean cake powder, silkworm chrysalis powder and the like; the inorganic nitrogen mainly comprises ammonium chloride, sodium nitrate, diammonium phosphate, etc. The organic nitrogen is best utilized. The mineral elements are mainly phosphorus, potassium, calcium, magnesium and the like. Inorganic salts are generally added to meet the requirements of cordyceps militaris on mineral elements. Vitamin Cordyceps sinensis mycelia cannot synthesize necessary vitamins, and VB1 is added properly to facilitate growth and development of strains.
In an alternative embodiment, the temperature is regulated during different stages of growth and development of Cordyceps militaris. The Cordyceps militaris strain should be propagated once and once regularly in production. Selecting high-yield, high-quality and early-matured cordyceps militaris entity, sterilizing the surface of the cordyceps militaris entity by using 0.1% mercuric chloride solution or 75% alcohol, cleaning the surface liquid medicine by using sterile water, suspending the cordyceps militaris entity above a container containing a comprehensive culture medium, standing and culturing at 21 ℃, and when a starburst cordyceps militaris colony appears on the surface of the culture medium, picking single or multiple colonies in an inoculation box and culturing in a test tube slant culture medium. Purifying after the Cordyceps militaris mycelium is fully distributed on the inclined plane. After the obtained sporophore is subjected to a grass-yielding comparison test, selecting high-quality cordyceps sporophore, performing tissue separation again, and screening to obtain the plant seed for transfer expansion cultivation. This is a well-known technique and will not be described in detail herein.
Referring to fig. 1 and 2, in this embodiment, the culture assembly 11 includes a culture bottom plate 111, filtering holes 112, a surrounding plate 113, a rotating shaft 114 and a movable shell 115, the surface of the culture bottom plate 111 is provided with the filtering holes 112 in an array, surrounding plates 113 are arranged around the top of the culture bottom plate 111, the rotating shaft 114 is arranged at two sides of the outer wall of the surrounding plate 113, the movable shell 115 is arranged at one side of the rotating shaft 114, and the movable shell 115 is sleeved outside the culture bottom plate 111.
Referring to fig. 1 and 3, in this embodiment, the soaking assembly 12 further includes a valve 124, a clamping plate 125, a tension spring 126 and a connecting block 127, the soaking tank 121 is detachably disposed on one side of the top of the culture bottom plate 111, the inlet end of the water outlet pipe 123 is connected to the soaking tank 121, the outlet end of the water outlet pipe 123 is provided with the valve 124, the valve 124 is used for enabling the liquid inside the soaking tank 121 to flow unidirectionally from the inlet end to the outlet end of the water outlet pipe 123, one side of the water outlet pipe 123 away from the soaking tank 121 is provided with the clamping plate 125, the number of the clamping plates 125 is two, the tension spring 126 is disposed between the two clamping plates 125, and the connecting block 127 is disposed on one side of the soaking tank 121 close to the coaming 113.
Importantly, one clamping plate 125 is fixed to the side of the outlet pipe 123 remote from the soaking tank 121, and the other clamping plate 125 is movably arranged by a tension spring 126.
Referring to fig. 1 and 4, in this embodiment, the detecting assembly 13 includes a collecting box 131, a display screen 132, a weight detector 133, an outlet channel 134, a baffle 135 and a sampling box 136, the collecting box 131 is disposed at the bottom of the culture bottom plate 111, an inlet end of the collecting box 131 is communicated with an outlet end of the filtering hole 112, one side of an outer surface of the collecting box 131 is provided with the display screen 132, the bottom of an inner wall of the collecting box 131 is obliquely disposed, the bottom of the inner wall of the collecting box 131 is provided with the weight detector 133, the other side of the outer surface of the collecting box 131 is provided with the outlet channel 134, one end of the outlet channel 134 is movably provided with the baffle 135, and an outlet end of the outlet channel 134 is provided with the sampling box 136.
Therefore, by environmental preparation in the cultivation method, the defecation amount of the larvae can be monitored in time by the weight detector 133 so that the defecation amount of the larvae and the cultivation days are in a normal proportion, and simultaneously, the defecation of the larvae is extracted so that the chlorophyll content in the defecation of the larvae is in a normal value, so as to detect the absorption degree of the larvae to the nutrient solution and the flavoring agent. When the defecation amount of the larvae is too high or too low, the state of the larvae is checked in time to prevent diseases and the like, alternatively, the weight detector 133 may be provided with an alarm device, and when the set defecation amount is out of or below a preset range, an alarm notifying personnel to check occurs. When the chlorophyll content in the excretions of the larvae is abnormal, the nutrient solution and flavoring agent content in the soaking tank 121 is checked and adjusted in time.
Referring to fig. 1, in this embodiment, the pupa formation assembly 14 includes a pupa formation outer box 141, a pupa formation inner box 142, and a container 143, wherein the pupa formation outer box 141 is disposed on the other side of the top of the culture bottom plate 111, the pupa formation inner box 142 is disposed inside the pupa formation outer box 141, and the container 143 is disposed on the surface of the pupa formation inner box 142 in an array.
In particular, in the preparation of the material, the absorption liquid comprises a nutrient solution and a flavoring agent,
the nutrient solution comprises 65-75 parts by weight of water, glucose, sucrose, maltose, starch and pectin: 10 to 15: 2-10: 2-10: 3 to 12:0.2 to 1;
the flavoring agent comprises salty agent, sweetener, flavoring agent, sour agent and Xin Xiangji, wherein the flavoring agent comprises sodium glutamate and inosinic acid, and the ratio of the sodium glutamate to the inosinic acid is 5-20: 1, a step of;
the cultured leaf is one or more of green grass, mulberry leaf, locust tree leaf, banyan leaf, camphor leaf, willow leaf and tender leaf;
the stem end of the culture leaf in the preparation of the material is inserted into the water outlet pipe 123, and the leaf end of the culture leaf is placed on the culture bottom plate 111.
Specifically, the qualification standard of the larvae in the environmental preparation is as follows: the larvae had no spots on the outer surface, the length of the larvae was 5 to 7cm, and the ratio of defecation amount to days of culture was 0.1g:1day, chlorophyll content in the stool is less than 15%.
In an alternative embodiment, the flavor Xin Xiangji is added with cruciferous phytochemicals, and Xin Xiangji with cruciferous phytochemicals is added into the soaking box 121, so that the leaf ends of the culture leaves absorb the cruciferous phytochemicals, after detecting that the chlorophyll content in the excrement of the larvae is lower than 15%, the strain of the cordyceps militaris is infected after the larvae are in a pupa state, the mature cordyceps militaris can have the cruciferous phytochemicals, cordycepin, cordycepic acid, cordyceps polysaccharide, ergosterol, adenosine, superoxide dismutase, flavonoids, carotene, various trace elements and the like, so that the cordyceps militaris has strong radish flavor, and the taste of the cordyceps militaris is changed; in another alternative embodiment, the seasoning Xin Xiangji is added with limonin, and Xin Xiangji with limonin is added into the soaking box 121, so that the leaf end of the cultivated leaf absorbs the limonin, after detecting that the chlorophyll content in the larva is lower than 15%, the mature cordyceps militaris can have limonin, cordycepin, cordycepic acid, cordyceps polysaccharide, ergosterol, adenosine, superoxide dismutase, flavonoids, carotene and various trace elements, etc. after the cordyceps militaris is subjected to pupation, the cordyceps militaris has a lighter and lighter lemon flavor.
It is worth noting that fruit flavoring agents can be used in the flavoring agent, and the fruit flavoring agents mainly comprise esters, aldehydes, ketones, acids, alcohols, volatile acids and the like, and the substances form fruit flavor according to certain concentration and proportion, so that the effect of improving the taste of cordyceps militaris is achieved, and the fruits only have unique fruit flavor by the compounds.
Specifically, the table is a fruit fragrance component detection table in cordyceps militaris;
specifically, the rotation method comprises the following steps: kaiser normalized maximum method difference
It is worth noting that according to the above theory and experimental analysis, after the larvae eat the culture leaves containing fruit flavor components, the fruit flavor substances in the culture leaves are absorbed, decomposed, metabolized and synthesized, so that part of the flavor substances remained in the larvae are gradually metabolized out of the body along with the time lapse, after the larvae become pupae, the step strain cultivation must be completed within 24 hours after the step strain breeding, so that the strain infects the insect pupae, and the cordyceps militaris with fruit flavor can be produced.
In another alternative embodiment, the flavoring agent is replaced with: flavoring agents of carvone, menthol, isoamyl acetate and anethole, and measuring the content of the corresponding substances in the larvae. The dose of all the substances in the infusion tank 121 is 100mg. The following figures show:
The concentration of various substances in the larvae at different times was recorded in this example, and it can be seen that after 2 hours, anethole reached a peak concentration, which was also the most intense flavor substance among the 3 substances that could be clearly detected.
The larvae were then collected over different time periods and analyzed by mass spectrometry. It follows that these substances appear in the larvae at the highest rate within 5 minutes after they ingest the cultured leaves. Carvone and anethole concentrations peak after 2 hours, after which they gradually decrease, while menthol stabilizes. The detected isoamyl acetate content was relatively low. After the flavoring agent is continuously added in the growth period of the larvae, the flavoring agents with the flavors of carvone, menthol, isoamyl acetate and anethole can continuously exist in the larvae and enter the pupae to be stored along with pupation.
Therefore, through the material preparation in the cultivation method, the nutrient solution and the flavoring agent in the soaking component 12 are absorbed by the stem end of the cultivation leaf, so that the nutrient solution and the flavoring agent are fused into the leaf end of the cultivation leaf, therefore, after the cultivation leaf containing personnel-controllable micro-elements is eaten by the larva, the cultivation leaf is absorbed by the larva in a conversion absorption mode, the nutrient solution and the flavoring agent are converted into micromolecular substances after being digested and absorbed by the stomach, the nutrient components and the needed flavoring agent contained in the larva are gradually improved and increased in a long-term adult period, the rejection rate of directly adding the nutrient solution and the flavoring agent into the cordyceps militaris is reduced, the nutrient solution and the flavoring agent are accumulated in the larva in a conversion absorption mode, the nutrient solution and the flavoring agent are absorbed by bacteria infected by the pupa in a hidden mode, the nutrient content of the cultivated cordyceps militaris is improved, the taste of the cordyceps militaris is controlled by the residual flavoring agent, the taste is convenient, more people adapt to the customized cordyceps militaris, and the audience and the prospect of the cordyceps militaris is expanded.
The invention also provides a method for purifying Cordyceps militaris,
the purification method is based on a purification device 2;
the purifying apparatus 2 includes: extraction assembly 21 and purification assembly 22, the output of extraction assembly 21 is connected to the input of purification assembly 22;
the purification method comprises the following steps:
crushing and drying: crushing the cultivated cordyceps militaris to a diameter of 150-200 um, and putting the cordyceps militaris into a baking oven with the temperature of 35-45 ℃ for baking;
filtering Cordyceps militaris powder: placing dried Cordyceps militaris into an extraction component 21, pouring distilled water, heating for 10min, extracting for 30min, and filtering with 0.45 μm microporous membrane;
purifying cordycepin: cordycepin is extracted through the purification component 22, the extraction temperature is 100 ℃, and the feed-liquid ratio is 1:15.
referring to fig. 5, in this embodiment, the extracting unit 21 includes: including filter paper 211, extractor 212, collector 213 and oil bath 214, filter paper 211 sets up in the inside of extractor 212, alternatively, the extractor 212 can be the soxhlet extractor, and the bottom of extractor 212 is connected with collector 213, and the bottom of collector 213 is equipped with oil bath 214.
Referring to fig. 5, in this embodiment, the purifying component 22 includes: distiller 221, condenser pipe 222, recovery bottle 223 and magnetic stirrer 224, distiller 221 is used for connecting collector 213, and one side of distiller 221 is equipped with condenser pipe 222, and the one end of condenser pipe 222 is connected in recovery bottle 223, and the bottom of recovery bottle 223 is equipped with magnetic stirrer 224.
Specifically, during the crushing and drying steps, the collected Cordyceps militaris powder is placed in the filter paper 211, the filter paper 211 is located in the extractor 212, wherein the filter paper 211 is tightly attached to the wall of the extractor 212, and the extracting liquid medicine is filled in the collector 213, in this embodiment, the cordycepin of the Cordyceps militaris powder is easily dissolved in distilled water, therefore, in this embodiment, distilled water is filled in the collector 213 for extracting the cordycepin, the collector 213 is placed in the oil bath pan 214, the distilled water wets the filter paper 211, the condensed water is started, the temperature is set in the oil bath pan 214, heating is started, and along with the rise of the temperature, the cordycepin of the Cordyceps militaris powder is dissolved in the distilled water, alternatively, siphoning can be performed for a plurality of times to enable the cordycepin of the Cordyceps militaris powder to be extracted as much as possible, and the mixed liquid of the cordycepin and the distilled water can be collected in the collector 213 for about 3 min. Alternatively, the supernatant may be centrifuged at 3000r/min for 10 minutes, the supernatant is taken out to a volume of about 100ml, and finally filtered by a 0.45 μm microporous filter membrane, at this time, the filtered collector 213 is mounted with a distiller 221, the collector 213 is placed in an oil bath 214, condensed water is turned on, the temperature of the oil bath 214 is set to start heating, the water temperature is maintained at 100 °, and since the boiling point of distilled water is 100 °, the distilled water is heated and evaporated, condensed by a condenser 222 and dropped into a recovery bottle 223, so that pure cordycepin is left in the recovery bottle 223. Since cordycepin in this embodiment is insoluble in water, a small amount of water is added to the recovery bottle 223 to precipitate cordycepin, the water is evaporated by heating with the magnetic stirrer 224, and the cordycepin is obtained in the recovery bottle 223 as the water is continuously evaporated.
Therefore, through the cooperation setting of extracting component 21 and purification subassembly 22, can carry out a lot of purification to the cordyceps militaris, further improve the purification degree of cordycepin.
Compared with the prior art, the technical scheme provided by the invention has at least the following beneficial effects:
in the above scheme, the materials in the cultivation method are prepared, so that the nutrient solution and the flavoring agent in the soaking component 12 are absorbed by the stem end of the cultivation leaf, and the nutrient solution and the flavoring agent are fused into the leaf end of the cultivation leaf, so that after the cultivation leaf containing personnel-controllable microelements is eaten by the larva, the larva can absorb the nutrient solution and the flavoring agent in a conversion absorption mode, the nutrient solution and the flavoring agent are converted into micromolecular substances after being digested and absorbed by the larva, the nutrient components and the required flavoring agent contained in the larva are gradually improved and increased in a long-term adult period, the rejection rate of directly adding the nutrient solution and the flavoring agent into the cordyceps militaris is reduced, the nutrient solution and the flavoring agent accumulated in the larva are absorbed in a conversion absorption mode, the nutrient solution and the flavoring agent are absorbed by the infected strain of the cordyceps militaris, the nutrient content of the cultivated cordyceps militaris is improved, the taste of the cordyceps militaris is controlled by the residual flavoring agent, the cordyceps militaris is customized, the cordyceps militaris is more suitable for the audience and the cordyceps militaris is expanded.
Through the combination setting of soaking case 121, can utilize stirring leaf 1214 of retaining space 1212 bottom to stir nutrient solution and the flavouring of injection constantly for nutrient solution and flavouring intensive mixing, and prevent the element precipitation of nutrient solution and flavouring, simultaneously, through guiding sponge layer 1215 and water storage sponge layer 1216, make the stem tip of cultivateing the leaf contact with water storage sponge layer 1216, make the stem tip of cultivateing the leaf receive the water storage sponge layer 1216 parcel of absorbed nutrient solution and flavouring, make the root and the surface of the stem tip of cultivateing the leaf all contact with water storage sponge layer 1216, the area of contact of the stem tip of cultivateing the leaf and water storage sponge layer 1216 has been improved, increase the absorption efficiency of the stem tip of cultivateing the leaf, and after the water level of nutrient solution and flavouring descends in retaining space 1212, through the conduction of guide sponge layer 1215, the stem tip of cultivateing the leaf still can stable absorption nutrient solution and flavouring, keep the sufficient of the leaf tip nutrition of cultivateing the leaf, do not need to inject the nutrient solution and flavouring constantly, the use is convenient.
Through the environmental preparation in the cultivation method, the defecation amount of the larvae can be monitored in time by using the weight detector 133, so that the defecation amount of the larvae and the cultivation days are in a normal proportion, and meanwhile, the defecation of the larvae is extracted, so that the chlorophyll content in the defecation of the larvae is in a normal value, and the absorption degree of the larvae to the nutrient solution and the flavoring agent is detected.
Through the cooperation setting of extracting assembly 21 and purification subassembly 22, can carry out the purification of a lot of to the cordyceps militaris, further improve the purification degree of cordycepin.
In conclusion, the invention has the advantages of reducing the rejection rate of directly adding nutrient solution and flavoring agent into the pupa, improving the nutrition content of the cultivated cordyceps militaris, controlling the taste of the cordyceps militaris, further improving the purity of cordycepin and the like.
The invention is intended to cover any alternatives, modifications, equivalents, and variations that fall within the spirit and scope of the invention. In the following description of preferred embodiments of the invention, specific details are set forth in order to provide a thorough understanding of the invention, and the invention will be fully understood to those skilled in the art without such details. In other instances, well-known methods, procedures, flows, components, circuits, and the like have not been described in detail so as not to unnecessarily obscure aspects of the present invention.
Those of ordinary skill in the art will appreciate that all or a portion of the steps in implementing the methods of the embodiments described above may be implemented by a program that instructs associated hardware, and the program may be stored on a computer readable storage medium, such as: ROM/RAM, magnetic disks, optical disks, etc.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (10)
1. A cultivation method of Cordyceps militaris is characterized in that,
the cultivation method is based on cultivation equipment;
the incubation apparatus includes: the device comprises a culture assembly, a soaking assembly, a detection assembly and a pupa formation assembly, wherein the soaking assembly is arranged on one side of the interior of the culture assembly, the detection assembly is arranged at the bottom of the culture assembly, the pupa formation assembly is arranged on the other side of the interior of the culture assembly, the soaking assembly comprises a soaking box, a liquid inlet and a water outlet pipe, the liquid inlet is arranged at the top of the soaking box, and the water outlet pipe is arranged on one side of the soaking box;
the soaking tank comprises a tank body, a water storage space, a motor, stirring blades, a guide sponge layer and a water storage sponge layer, wherein the water storage space is formed in the bottom of the inner wall of the tank body and is communicated with the liquid inlet, the motor is arranged at the bottom of the tank body, the stirring blades are arranged at the bottom of the water storage space and are connected with the motor, the top of the inner wall of the tank body is filled with the water storage sponge layer, the guide sponge layer is arranged between the water storage space and the water storage sponge layer, and the water storage sponge layer is communicated with the water outlet pipe;
The cultivation method comprises the following steps:
material preparation: placing the larvae into the culture assembly, adding absorption liquid into the soaking assembly, inserting stem ends of the culture leaves into the soaking assembly, placing leaf ends of the culture leaves into the culture assembly, and providing nutrients with the absorption liquid for the larvae;
preparing environment: detecting the state of the larvae through the detection assembly, keeping the increase of the defecation amount of the larvae stable, picking out unqualified larvae, and waiting for the qualified larvae to form pupae in the pupae formation assembly;
seed selection of strains: selecting excellent strains, sterilizing and extracting strains;
culturing strains: sterilizing the surface of the pupa, infecting the pupa by strain, and culturing the infected pupa in aseptic environment.
2. The method according to claim 1, wherein the culturing component comprises a culturing bottom plate, filtering holes, a surrounding plate, a rotating shaft and a movable shell, wherein the surface of the culturing bottom plate is provided with the filtering holes in an array, the surrounding plate is arranged on the periphery of the top of the culturing bottom plate, the rotating shaft is arranged on two sides of the outer wall of the surrounding plate, the movable shell is arranged on one side of the rotating shaft, and the movable shell is sleeved on the culturing bottom plate.
3. The method according to claim 2, wherein the soaking assembly further comprises a valve, clamping plates, tension springs and connecting blocks, the soaking box is detachably arranged on one side of the top of the culture bottom plate, the inlet end of the water outlet pipe is communicated with the soaking box, the outlet end of the water outlet pipe is provided with a valve, the valve is used for enabling liquid in the soaking box to flow unidirectionally from the inlet end to the outlet end of the water outlet pipe, the clamping plates are arranged on one side of the water outlet pipe away from the soaking box, the number of the clamping plates is two, the tension springs are arranged between the two clamping plates, and the connecting blocks are arranged on one side of the soaking box close to the coaming.
4. The cordyceps militaris cultivating and purifying method as in claim 3, wherein the detecting assembly comprises a collecting box, a display screen, a weight detector, an outlet channel, a baffle and a sampling box, the collecting box is arranged at the bottom of the cultivating base plate, the inlet end of the collecting box is communicated with the outlet end of the filtering hole, the display screen is arranged on one side of the outer surface of the collecting box, the bottom of the inner wall of the collecting box is obliquely arranged, the weight detector is arranged at the bottom of the inner wall of the collecting box, the outlet channel is arranged on the other side of the outer surface of the collecting box, the baffle is movably arranged at one end of the outlet channel, and the sampling box is arranged at the outlet end of the outlet channel.
5. The method according to claim 4, wherein the pupa formation component comprises an outer pupa formation box, an inner pupa formation box and a containing box, the outer pupa formation box is arranged on the other side of the top of the culture bottom plate, the inner pupa formation box is arranged in the outer pupa formation box, and the containing boxes are arranged on the surface of the inner pupa formation box in an array.
6. The method for cultivating and purifying Cordyceps militaris according to claim 5, wherein in the preparation of the material, the absorption liquid comprises a nutrient solution and a flavoring agent,
the nutrient solution comprises 65-75 parts by weight of water, glucose, sucrose, maltose, starch and pectin: 10 to 15: 2-10: 2-10: 3 to 12:0.2 to 1;
the flavoring agent comprises salty agent, sweetener, flavoring agent, sour agent and Xin Xiangji, wherein the flavoring agent comprises sodium glutamate and inosinic acid, and the ratio of the sodium glutamate to the inosinic acid is 5-20: 1, a step of;
the cultured leaf is one or more of green grass, mulberry leaf, locust tree leaf, banyan leaf, camphor leaf, willow leaf and tender leaf;
the stem end of the culture leaf is inserted into the water outlet pipe in the preparation of the material, and the leaf end of the culture leaf is placed on the culture bottom plate.
7. The method for cultivating and purifying Cordyceps militaris according to claim 6, wherein the qualification criteria of the larvae in the environmental preparation are: the external surface of the larva is free from spots, the length of the larva is 5-7 cm, and the ratio of defecation amount to culture days is 0.1g:1day, chlorophyll content in the stool is less than 15%.
8. A method for purifying Cordyceps militaris is characterized in that,
the purification method is based on a purification device;
the purification apparatus includes: the output end of the extraction component is connected with the input end of the purification component;
the purification method comprises the following steps:
crushing and drying: crushing the cultivated cordyceps militaris to a diameter of 150-200 um, and putting the cordyceps militaris into a baking oven with the temperature of 35-45 ℃ for baking;
filtering Cordyceps militaris powder: placing the dried Cordyceps militaris into the extraction component, pouring distilled water, heating for 10min, extracting for 30min, and filtering with 0.45 μm microporous membrane;
purifying cordycepin: extracting cordycepin by the purification component, wherein the extraction temperature is 100 ℃, and the feed-liquid ratio is 1:15.
9. the method for cultivating and purifying Cordyceps militaris according to claim 8, wherein the extraction module comprises: the device comprises a filter paper piece, an extractor, a collector and an oil bath pot, wherein the filter paper piece is arranged in the extractor, the collector is connected to the bottom of the extractor, and the oil bath pot is arranged at the bottom of the collector.
10. The method of cultivating and purifying Cordyceps militaris according to claim 9, wherein the purifying assembly comprises: distiller, condenser pipe, recovery bottle and magnetic stirrer, the distiller is used for connecting the collector, one side of distiller is equipped with the condenser pipe, the one end of condenser pipe is connected in the recovery bottle, the bottom of recovery bottle is equipped with magnetic stirrer.
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